Calcium and trace mineral supplements comprising estrogen

ABSTRACT

Nutritional mineral supplements comprising calcium citrate malate and salts of manganese, copper and zinc are disclosed. These supplements, which provide at least 25% RDA of the minerals, are used in addition to the normal diet. These supplements are useful for increasing bone growth and for treating age-related bone loss in humans and animals.

This is a division of application Ser. No. 07/562,773, filed on Aug. 6,1990 U.S. Pat. No. 5,151,274.

TECHNICAL FIELD

The present invention relates to nutritional improvements in calciumsupplements containing trace minerals, in particular copper, manganeseand zinc. These supplements are useful for increasing bone growth andtreating age-related bone loss. They can be used in conjunction withfoods and beverages or taken as an oral solid or liquid supplement. Theinvention also relates to a method of building bone or treating boneloss in osteoporosis patients and post-menopausal women.

BACKGROUND OF THE INVENTION

Vitamin and mineral supplements for human and veterinary use arecommonplace. Some diets, heavy physical exercise and disease conditionsmay require the intake of considerable quantities of minerals apart fromthose generally obtained through what otherwise would be considered anormal diet. Calcium and trace mineral supplementation is importantprimarily for those who have inadequate diets, including growingchildren. Older adults have an additional need for calcium to helpprevent the bone loss which occurs as a normal consequence of the agingprocess. In particular, postmenopausal women need additional calcium dueto hormonal changes which can accelerate the bone loss rate leading to afurther diminishment in bone mass.

The trace minerals which affect bone growth are copper, zinc andmanganese. Supplementation of the diet with these minerals along with ahighly bioavailable source of calcium is highly desirable. Commerciallyavailable mineral supplements are useful in many circumstances whereincreased mineral intake is desirable. Most of these multi-vitamin andmulti-mineral tablets are low in calcium, requiring separatesupplementation with calcium sources. In addition, not all calciumsources are equal in terms of bioavailability and absorption. It wouldbe more convenient if all of the minerals could be administeredconjointly in a convenient and/or pleasant tasting form which would notrequire extra attention, planning and implementation by the user. Thiscould be done in the form of foods and beverages as well as in the formof tablets.

There are well-recognized problems associated with adding both mineraland calcium supplements to foods and beverages. Some of these are taste;calcium tends to be chalky in flavor. In addition, the solubility ofmany calcium sources prevents them from being added to many beverages.Others are interactions of the minerals with the food or beverage whichaffects the stability and/or the bioavailabilty of the product. Thisinvention provides a means for making such product.

This invention also relates to methods of building bone in humans andother animals, i.e., for the treatment of age-related bone loss andrelated disorders. In particular, this invention relates to such methodsof treatment by administration of certain calcium salts and theminerals, copper, zinc and manganese.

Calcium is the fifth most abundant element in the human body. It playsan important role in many physiological processes, including nerve andmuscle functions. Not surprisingly, nutritional and metabolicdeficiencies of calcium can have broad-ranging adverse effects. Sinceabout 98% to 99% of the body's calcium is found in bone tissues, many ofthese adverse effects are manifested through deficiencies in thestructure, function and integrity of the skeletal system.

The most common metabolic bone disorder is osteoporosis. Osteoporosiscan be generally defined as the reduction in the quantity of bone,either from the reduction in bone formation or the acceleration of boneresorption, in either event the result is a decrease in the amount ofskeletal tissue. In general, there are two types of osteoporosis:primary and secondary. "Secondary osteoporosis" is the result of anidentifiable disease process or agent. However, approximately 90% of allosteoporosis cases are idiopathic "primary osteoporosis". Such primaryosteoporosis includes postmenopausal osteoporosis, age-associatedosteoporosis (affecting a majority of individuals over the age of 70 to80), and idiopathic osteoporosis affecting middle-aged and younger menand women.

For some osteoporotic individuals the loss of bone tissue issufficiently great so as to cause mechanical failure of the bonestructure. Bone fractures often occur, for example, in the wrist, hipand spine of women suffering from postmenopausal osteoporosis. Kyphosis(abnormally increased curvature of the thoracic spine) may also result.

The mechanism of bone loss in osteoporotics is believed to involve animbalance in the process of "bone remodeling". Bone remodeling occursthroughout life, renewing the skeleton and maintaining the strength ofbone. Two reactions are involved, bone loss or resorption and bonegrowth or accretion. This remodeling occurs in a series of discretepockets of activity in the bone, called "osteoclasts" and "osteoblasts".Osteoclasts (bone dissolving or resorbing cells) are responsible for theresorption of a portion of bone within the bone matrix, during theresorption process. After resorption, the osteoclasts are followed bythe appearance of osteoblasts (bone forming cells), which then refillthe resorbed portion with new bone.

In young healthy adults, the rate at which the osteoclasts andosteoblasts are formed maintains a balance of bone resorption and boneformation. However, as normal consequency of aging an imbalance in thisremodeling process develops, resulting in loss of bone at a rate fasterthan the accretion of bone. As imbalance continues over time thereduction in bone mass and thus bone strength leads to fractures.

Many compositions and methods are described in the medical literaturefor the "treatment" of osteoporosis. See, for example, R. C. Haynes, Jr.et al., "Agents affecting Calcification", The Pharmacological Basis ofTherapeutics, 7th Edition (A. G. Gilman, L. S. Goodman et al., Editors,1985); and G. D. Whedon et al., "An Analysis of Current Concepts andResearch Interest in Osteoporosis", Current Advances in Skeletogenesis(A. Ornoy et al., Editors, 1985). Estrogen is often used to affect themetabolism of calcium by influencing the osteoblast cells. Treatmentsusing fluoride have also been described. However, the utility of suchagents may be limited, because of possible adverse side effects. See W.A. Peck, et al., Physician's Resource Manual on Osteoporosis (1987),published by the National Osteoporosis Foundation.

Nutritional therapies for osteoporosis have also been proposed. Manycalcium-containing compounds and compositions have been described foruse as nutritional supplements. Many commercial preparations are alsoavailable, typically containing calcium carbonate or calcium phosphate.Other calcium salts have also been described for use in calciumsupplements, including calcium lactate, calcium citrate and calciumgluconate.

U.S. Pat. No. 3,949,098 issued Bangert (assigned Nabisco, 1976)describes a nutritious orange drink concentrate that contains wheyprotein. The patent suggests the addition of minor amounts of vitaminsand other nutrients which include various cupric salts, manganese salts,zinc salts, as well as calcium salts.

German OLS 2,845,570 issued to E.R.E. (Europe RepresentationEstablishment, 1980) describes a honey containing composition. Honeycontains low levels of calcium, manganese, copper as well as traceamounts of magnesium, iron, phosphorous, silicon and nickel. The valueof honey as a medicant is undisputed according to this patentapplication. This application claims a honey containing composition withlevarotatory ascorbic acid and citric acid. This patent has issued asU.S. Pat. No. 4,243,794 (1981).

U.S. Pat. No. 4,497,800 issued to Larsen et al (assigned Mead Johnson &Company, 1985) describes a nutritionally complete ready-to-use liquiddiet for providing total patient nourishment. The diet contains freeamino acids and small peptides, a carbohydrate source, and nutritionallysignificant amounts of all essential vitamins and minerals, andstabilizers. The minerals include calcium, copper, zinc and manganese,among others. Most of these minerals are given as the gluconate salt.

"Effects of calcium carbonate in hydroxyapatite on zinc and ironretention in postmenopausal women", Dawson-Hughes, Seligson and Hughes,American Journal of Clinical Nutrition, 44, 83-88 (1986) describes theeffect of calcium carbonate on whole-body retention of zinc and iron inthirteen healthy post menopausal women. The test meal, including bothdry food and a formulated beverage, included calcium, copper and zinc ata level of one-third the usual daily requirement. These are levelsnormally found in human diets.

U.S. Pat. No. 3,992,555 issued to Kovacs (assigned Vitamins, Inc., 1976)describes food supplements prepared by mixing assimilable ironcompounds, vitamins and minerals with a heated edible fat carrier. Theminerals include calcium, zinc, copper, and manganese among others.

U.S. Pat. No. 3,950,547 issued to Lamar et al (assigned Syntex Inc,1976) describes a dietary composition containing peptides and/or aminoacids, lipids and carbohydrates in an aqueous emulsion. Suitableminerals for adding at low levels include among others calcium, copperand zinc.

U.S. Pat. No. 4,107,346 issued to Kravitz (1978) describes a dietarysalt composition for use as a replacement for salt in foods. The role ofcopper is described as a component of several enzymes essential fornutrition. The patent further discloses that "spontaneous fractures arecommon in animals feeding off copper deficient soils or who are givenartificially depleted copper diets". (column 4, lines 20-30) Zinc isdescribed as helping in growth, wound healing and improving taste andsmell. Clinical symptoms of manganese deficiency have not been observedin man. However, there is no question that manganese is essential forhuman nutrition. Main manifestations of its deficiency are impairedgrowth and skeletal abnormalities. Calcium is described as beingnecessary for blood coagulation, for calcium retention, and forrelieving the symptoms of osteoporosis. Examples of salts that containthese four trace minerals are disclosed.

U.S. Pat. No. 4,070,488 issued to Davis (unassigned, 1978) discloses ahighly stabilized balanced nutritive composition useful in supplementingthe diet of humans and/or animals. This composition contains gelatin.The patent discloses that the sulfhydryl groups of the gelatin canrender copper inactive toward ascorbic acid.

U.S. Pat. No. 4,214,996 issued to Buddemeyer et al (R.G.B. Laboratories,1980) discloses mineral compositions which are very soluble. Thesecompositions contain calcium, phosphorus, zinc, as well as manganese.Not all of the compositions that are described contain all fourelements.

U.S. Pat. No. 4,351,735 to Buddemeyer et al (R.G.B. Laboratories, 1982)is related to the '996 patent.

"Nutrients and Nutrition of Citrus Fruits," Citrus Nutrition andQuality, Ting, (American Chemical Society, 1980) discloses the presenceof certain trace minerals in orange juice. These include copper, zinc,iron and manganese. Calcium and magnesium are the two major divalentcations in orange juice. The levels of all the minerals are low.

Hungerford et al, "Interaction of pH and ascorbate in intestinal ironabsorption," (1983) describes the iron absorption from various foodmaterials. The diet which was low in iron also contained calciumcarbonate, manganese sulfate and copper sulfate among others.

U.S. Pat. No. 4,419,369 issued to Nichols et al (assigned Baylor Collegeof Medicine, 1983) describes an improved dietary protein mineral modulefor infants. An approximate analysis of the material shows the presenceof iron, zinc, copper and calcium.

The utility of these known supplements varies. Unlike agents (such asestrogen) which affect the metabolism of bone, calcium nutritionalsupplements have been thought to merely provide a source for calcium(which may or may not be properly absorbed and metabolized). See, forexample, B. Riis et al., "Does Calcium Supplementation PreventPostmenopausal Bone Loss?," New England J. of Medicine, 316, 173-177(1987); L. Nilas et al., "Calcium Supplementation and PostmenopausalBone Loss," British Medical Journal, 289, 1103-1106 (1984); and H.Spencer et al., "NIH Concensus Conference: Osteoporosis," Journal ofNutrition, 116, 316-319 (1986).

It has now been discovered, however, that the administration of mixturesof certain calcium salts, along with trace minerals, copper, zinc andmanganese are surprisingly effective for delaying age-related loss ofbone. In particular, as compared to nutritional regimens known in theart, these methods afford greater efficacy in the treatment ofage-related bone loss and related disorders.

It would be desirable, therefore, to have mixed calcium and mineralsupplements which are compatible and nutritionally available. It wouldalso be quite useful to have such supplements which could be added tofood and beverage compositions without undesirably affectingorganoleptic or aesthetic properties.

It is an object of the present invention to provide calcium mineralsupplements which provide bone growth and can be used to treatage-related bone loss or to correct the imbalance that occurs betweenbone formation and bone resorption.

It is a further object of this invention to provide foodstuffs,beverages and beverage concentrates which are supplemented with calciumand trace minerals.

These and other objects are secured herein, as will be seen from thefollowing disclosure.

SUMMARY OF THE INVENTION

The multimineral supplements employ specific calcium salts of mixturesof citric and malic acids. The copper, manganese and zinc salts aresalts of sulfate, nitrate and chloride and carboxylates, e.g.gluconates.

The present invention provides methods for building bone in a human orother animal subject, comprising administering to said subject a safeand effective amount of calcium citrate malate and of copper, zinc andmanganese salts. The calcium citrate malate comprises a complex or amixture of calcium salts having a ratio of moles citrate to moles malateof from about 1:0.16 to about 1:13.5. The calcium citrate malate ispreferably administered in an oral dosage form, containingpharmaceutically-acceptable carriers and excipients.

All ratios, proportions and percentages herein are by weight, unlessotherwise specified. All weights of the minerals are on an elementalbasis unless otherwise specified.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to stable mineral supplements andsupplemented foods and beverages including dry beverage mixes and to amethod of building bone.

As used herein, the term "comprising" means various components can beconjointly employed in the mineral supplements, foods and beverages ofthe present invention. Accordingly, the terms "consisting essentiallyof" and "consisting of" are embodied in the term comprising.

By "nutritional" or "nutritionally-supplemental amount" herein is meantthat the mineral sources used in the practice of this invention providea nourishing amount of said minerals. This supplemental amount willcomprise at least 25% of the Recommended Dietary Allowance (RDA) of thedaily intake of calcium, copper, manganese and zinc. Preferably, atleast 50% of the Recommended Dietary Allowance (RDA) will be provided.The RDA for minerals is as defined in The United States of America (seeRecommended Daily Dietary Allowance-Food and Nutrition Board, NationalAcademy of Sciences-National Research Council). This is supplemental orin addition to the amount found in the diet.

As used herein, the term "flavors" includes both fruit and botanicalflavors.

As used herein the term "sweeteners" includes sugars, for example,glucose, sucrose, and fructose. Sugars also include high fructose cornsyrup solids, invert sugar, sugar alcohols, including sorbitol, andmixtures thereof. Artificial sweeteners are also included in the termsweetener.

As used herein, the term "trace minerals" means copper, manganese andzinc. These minerals play an important role in nutrition, but arerequired in only small or trace amounts in the diet. All three of theseminerals are important enzymatic cofactors which are essential indevelopment of bone in animals and humans. The trace minerals herein areadministered in the form of pharmaceutically acceptable salts.

The "carboxylate counterion" used in the preparation of the preferredmineral salts herein can be any ingestible carboxylate species. However,some judgement must be made with regard to flavor contribution. Forexample, citrate, malate and ascorbate yield ingestible complexes whoseflavors are judged to be quite acceptable, particularly in fruit juicebeverages. Tartaric acid is acceptable, particularly in grape juicebeverages, as is lactic acid. Longer-chain fatty acids may be used insolid mineral supplements, but can affect flavor and water solubility.For essentially all purposes, the malate (preferred), gluconate, citrateand ascorbate moieties suffice, although others can be selected,according to the desires of the formulator.

The counterion for the trace minerals can also be phosphate, chloride,sulfate, nitrate or the like. However, such counterions can undesirablyinteract with calcium ions, especially in beverages. In highconcentrations, these counterions, particularly chloride and sulfate,may contribute an undesirable flavor note. Accordingly, the carboxylatecounterions noted above are preferred herein.

The methods of the present invention comprise the administration ofcalcium citrate malate to a human or other animal subject along with thetrace mineral salts. Specific compounds and compositions to be used inthese processes must, accordingly, be pharmaceutically acceptable. Asused herein, a "pharmaceutically acceptable" component is one that issuitable for use with humans and/or animals without undue adverse sideeffects (such as toxicity, irritation, and allergic response)commensurate with a reasonable benefit/risk ratio. Further, as usedherein, the term "safe and effective amount" refers to the quantity of acomponent which is sufficient to yield a desired therapeutic responsewithout undue adverse side effects (such as toxicity, irritation, orallergic response) commensurate with a reasonable benefit/risk ratiowhen used in the manner of this invention. The specific "safe andeffective amount" will, obviously, vary with such factors as theparticular condition being treated, the physical condition of thepatient, the duration of the treatment, the nature of concurrent therapy(if any), and the specific formulations employed.

Trace Mineral Component

In supplements of the type disclosed herein, the nutritionallysupplemental amount for the minerals will generally comprise more than50% of the RDA and preferably 80%-100% RDA, most preferably 100% of theRDA, per unit portion of the finished supplement. Of course, it isrecognized that the preferred daily intake of any mineral may vary withthe user.

In general, the RDA (calcium) will range from 360 mg per 6 Kg forinfants to 800 mg/54-58 Kg female, depending somewhat on age. Moreover,it can be difficult to supplement beverages with more than 20-30% RDA ofcalcium (based per serving) without encountering precipitation and/ororganoleptic problems. However, this level of supplementation isequivalent to cow's milk in calcium value, and is therefore acceptable.

The estimated safe and adequate daily intake for zinc is 15 milligrams(mg) per day for males and 12 mg per day for females. There is nospecific RDA for manganese and copper. A safe and adequate range hasbeen established as 2 to 5 mg for manganese and for copper, the range is1.5 mg to 3 mg per day.

Any soluble salt of the trace minerals can be used, for example, zincchloride, zinc sulfate, manganese sulfate, manganese gluconate, coppersulfate and copper gluconate are useful. A nutritionally supplementalamount of these minerals is used. However, the particular salt used andthe level will depend upon their interaction with other supplementingredients.

Inorganic anions which are useful for making the trace mineral salts aresulfate, nitrate, phosphate, hydrogen phosphate and carbonate.

It is essential to this supplementation that the calcium salts besoluble in the stomach. This solubilization aids in making the calciummore readily bioavailable. It is equally important that the traceminerals be solubilized and absorbed by the stomach and or intestine.Therefore the choice of calcium and mineral salts depends upon theinteraction of the salts in acid (stomach pH) solutions or basic(intestinal pH) solutions.

Solubility also plays an important role in the preparation of foods andbeverages containing these supplements.

Calcium Citrate Malate Compositions:

The methods of this invention involve administration of a mixture ofcalcium salts, herein "calcium citrate malate," comprising calcium saltsof citric acid and malic acid. The calcium citrate malate may consist ofa mixture of calcium citrate and calcium malate, a complex of calciumcontaining citrate and malate ligands, a mixture of a calcium salt withcitric acid and malic acid, or combinations thereof. (Mixtures of acalcium salt and citric and malic acids may be used to form calciumcitrate malate in situ, in a liquid dose form, or in the acidenvironment of the stomach of the subject to whom the mixture isadministered.) Preferred are calcium citrate malate mixtures made byadding calcium carbonate, calcium hydroxide or other suitable source toa mixture of citric and malic acid.

The molar ratio of citrate:malate is from about 1:0.16 to about 1:13.5,preferably from about 1:0.5 to about 1:4.5, more preferably from about1:0.75 to about 1:3. The ratio of moles calcium:total molescitrate:total moles of malate is from about 2:1:1 to about 6:3:4,preferably from about 4:2:3 to about 6:3:4. The calcium citrate malatemay contain other acid anions in addition to citrate and malate. Suchanions may include, for example, carbonate, hydroxide, and mixturesthereof depending on the calcium source.

Preferably, the calcium citrate malate is neutral, comprised entirely ofcitrate and malate anions. Thus, preferably, the equivalents of calcium(2×moles calcium) is about equal to the total number of equivalents ofcitrate (3×moles citrate) plus malate (2×moles malate). A preferredcalcium citrate malate has a calcium:citrate:malate molar composision ofabout 6:2:3.

The calcium citrate malate and trace mineral salts for use in themethods of this invention may be provided in solid or liquid dosageforms. Calcium citrate malate for use in solid forms may be made, forexample, by first dissolving citric acid and malic acid, in the desiredmolar ratio, in water. Calcium carbonate may then be added to thesolution, in such amount that the ratio of moles calcium to molescitrate and moles malate is as desired. Carbon dioxide will be evolved.The solution may then be dried (as by freeze drying or oven drying attemperatures below 100° C.) to obtain the calcium citrate malate.Methods for making calcium citrate malate are described in the followingdocuments: Co-pending application of Fox et al, Ser. No. 07/537313 filedJun. 14, 1990; Japanese Patent Specification SHO 56-97248, Kawai,published Aug. 5, 1981; and in U.S. Pat. No. 4,722,847 issued to Heckert(1988).

Flavor Component

The flavor component of the present invention contains flavors selectedfrom natural flavors, botanical flavors and mixtures thereof. The term"fruit flavors" refers to those flavors derived from the ediblereproductive part of a seed plant, especially one having a sweet pulpassociated with the seed. Also included within the term "fruit flavor"are synthetically prepared flavors made to simulate fruit flavorsderived from natural sources.

The term "botanical flavor" refers to flavors derived from parts of aplant other than the fruit; i.e. derived from bean, nuts, bark, rootsand leaves. Also included within the term "botanical flavor" aresynthetically prepared flavors made to simulate botanical flavorsderived from natural sources. Examples of such flavors include cocoa,chocolate, vanilla, coffee, kola, tea, and the like. Botanical flavorscan be derived from natural sources such as essential oils and extracts,or can be synthetically prepared.

The particular amount of the flavor component effective for impartingflavor characteristics to the supplements and food or beverage mixes ofthe present invention ("flavor enhancing") can depend upon the flavor(s)selected, the flavor impression desired, and the form of the flavorcomponent. The flavor component can comprise at least 0.05% by weight ofthe beverage composition and preferably from 0.05% to about 10%. Theamount of flavor added to the food, beverage or supplement is within theskill of one in the art and depends on the flavor intensity desired.

For chocolate or cocoa, the amount of flavor added is from about 0.05%to about 20%. Lower levels of artificial or synthetic chocolate flavorsare used than for cocoa itself.

The beverages can be flavored with fruit or other botanical flavors, eg., vanilla, strawberry, cherry, pineapple, banana, and mixturesthereof.

Sweetener Component

The sweetener composition is usually a monosaccharide or a disaccharide.These include sucrose, fructose, dextrose, maltose and lactose. Othercarbohydrates can be used if less sweetness is desired. Mixtures ofthese sugars can be used.

In addition to sugar of the present invention can contain other naturalor artificial sweeteners. Other suitable sweeteners include saccharin,cyclamates, acetosulfam, L-aspartyl-L-phenylalanine lower alkyl estersweeteners (e.g. aspartame), L-aspartyl-D-alanine amides disclosed inU.S. Pat. No. 4,411,925 to Brennan et al., issued Oct. 23, 1983,L-aspartyl-D-serine amides disclosed in U.S. Pat. No. 4,399,163 atBrennan et al., issued Aug. 16, 1983,L-aspartyl-L-1-hydroxymethylalkaneamide sweeteners disclosed in U.S.Pat. No. 4,338,346 to Brand, issued Dec. 21, 1982,L-aspartyl-1-hydroxyethylalkaneamide sweeteners disclosed in U.S. Pat.No. 4,423,029 to Rizzi, issued Dec. 27, 1983, L-aspartyl-D-phenylglycineester and amide sweeteners disclosed in European Patent Application168,112 to J. M. Janusz, published Jan. 15, 1986, and the like. Aparticularly preferred sweetener is aspartame.

The amount of the sweetener effective in the food, beverage, mixes orsupplements of the invention depends upon the particular sweetener usedand the sweetness intensity desired. For noncaloric sweeteners, thisamount varies depending upon the sweetness intensity of the particularsweetener. For sugar (i.e., sucrose), this amount can be from 10% to 85%(typically from 55% to 70%) by weight. In determining the amount ofsugar, any sugar or other sweetener present in the flavor component isalso included. Low-calorie sweetener combinations containing anoncaloric sweetener such as aspartame and a sugar, such as corn syrupsolids, or sugar alcohols can also be used in beverage mixes. Ingeneral, the amount of sweetener will be from about 0.5% to about 85%.

Other Ingredients

Other minor ingredients are frequently included in supplements, foodsand beverages. Such ingredients include preservatives such as benzoicacid and salts thereof, sulfur dioxide, butylated hydroxyanisole,butylated hydroxytoluene, etc. Also, typically included are colorsderived either from natural sources or synthetically prepared.

Salt, e.g. sodium chloride, and other flavor enhancers can be used toimprove the flavor of the food, beverage or supplement.

Emulsifiers can also be included. Any food grade emulsifier can be used.Lecithin is a preferred emulsifier. Other edible emulsifiers includemono- and diglycerides of long chain fatty acids, preferably saturatedfatty acids, and most preferably, stearic and palmitic acid mono- anddiglycerides. Propylene glycol esters are also useful in beverage mixes.

Fats or oils can also be added to supplements or foods to make them morepalatable.

Supplement Forms

Solid forms include tablets, capsules, granules and bulk powders.Tablets may contain suitable binders, lubricants, diluents,disintegrating agents, coloring agents, flavoring agents, flow-inducingagents and melting agents. Liquid oral dosage forms include aqueoussolutions, emulsions, suspensions, solutions and/or suspensionsreconstituted from non-effervescent granules and effervescentpreparations reconstituted from effervescent granules. Such liquid oraldosage forms may contain, for example, suitable solvents, preservatives,emulsifying agents, suspending agents, diluents, sweeteners, meltingagents, and coloring and flavoring agents. A preferred liquid dosageform contains calcium citrate malate and the trace minerals in ajuice-containing beverage or other beverage.

The trace minerals and calcium citrate malate can be coadministered inone tablet, liquid, food or beverage or they can be administeredseparately. A capsule containing the trace mineral salts and a secondtablet with the calcium citrate malate are easy to formulate and toswallow. A mineral supplement could also be coadministered with acalcium beverage.

Specific examples of pharmaceutically acceptable carriers and excipientsthat may be used to formulate oral dosage forms of the present inventionare described in U.S. Pat. No. 3,903,297, Robert, issued Sep. 2, 1975.Techniques and compositions for making dosage forms useful in themethods of this invention are described in the following references; 7Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors,1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981);and Ansel, Introduction to Pharmaceutical Dosage Forms 2nd Edition.(1976).

Method of Building Bone

Various oral dosage forms of calcium citrate malate and trace mineralsmay be used in the present invention. Such dosage forms comprise a safeand effective amount of calcium citrate malate, trace minerals and apharmaceutically acceptable carrier. Preferably the pharmaceuticallyacceptable carrier is present at a level of from about 0.1% to about99%, preferably from about 0.1% to about 75%, by weight of thecomposition. Unit dosage forms (i.e., dosage forms containing an amountof calcium citrate malate suitable for administration in one singledose, according to sound medical practice) preferably contain from about100 mg to about 1000 mg, preferably from about 100 mg to about 500 mg,more preferably from about 200 mg to about 300 mg of calcium (on anelemental basis). Unit dosage forms of copper (on an elemental basis)contain 0.5 to 5 mg, preferably 0.5 to 4.0 mg, of manganese (on anelemental basis) contain 1 to 8 mg and preferably from 2 mg to 7 mg, andof zinc (on an elemental basis) contain 1.5 to 30 mg and preferably 7.5to 20 mg zinc.

Specifically, the present invention provides a method for building bonein a human or other animal subject, comprising administering to saidsubject a safe and effective amount of calcium citrate malate, andcopper, zinc and manganese for a period of time sufficient to achieve anincrease in the net skeletal mass of said subject. As used herein,"building bone" refers to a decrease in the net skeletal loss of bone ofthe subject treated and therefore a net skeletal increase in mass. Theslowing of the rate of bone loss and the increase in growth rate occursimultaneously so the net bone density may stay the same. The increasein mass may be at any skeletal site, including spine, hip, long bones ofarms or legs. Preferably, the net skeletal mass is increased by at leastabout 0.1%, more preferably at least about 1%.

The loss of bone is cumulative over a long period of time. Typically,lifetime loss in bone mass is about 35% in males and 50% in females.Thus, even though a net skeletal increase of as little as 0.5% in oneyear is not particularly critical, over 10 years this results in 5% morebone mass than would be present if bone loss continued at its usualrate.

"Administering" refers to any method which, in sound medical practice,delivers the calcium citrate malate and trace minerals used in thisinvention to the subject to be treated in such a manner so as to beeffective in the building of bone. Preferably, the calcium citratemalate is administered orally in 2 doses a day and the mineralsupplements are orally administered in one dose a day. The calciumcitrate malate and minerals could be mixed and administered in 2 or 3doses a day if desired.

Preferably, from about 175 milligrams to about 2000 milligrams ofcalcium (as elemental calcium) are administered to said subject, perday. More preferably, from about 250 milligrams to about 1500milligrams, most preferably from about 500 milligrams to about 1000milligrams, of calcium are administered, per day. The specific amount ofcalcium citrate malate to be administered depends upon the relativepercentage weight of calcium in the particular calcium citrate malateemployed. From 0.5 to 5.0 mg of copper, 1.5 to 30 mg of zinc and 1 to 8mg of manganese are administered. The preferred daily dose total is 1.0to 3.0 mg copper, 7.5 to 20 mg zinc and 2 to 7 mg manganese. The weightis for the metal ion and not for the salt. By way of example, 19.9 mg ofmanganese chloride tetrahydrate supplies 5.5 mg manganese on anelemental basis.

The specific period of time sufficient to achieve an increase in the netskeletal mass of the subject may depend on a variety of factors. Suchfactors include, for example, the specific mineral formulation employed,the amount of minerals administered, the age and sex of the subject, thespecific disorder to be treated, concomitant therapies employed (ifany), the general physical health of the subject (including the presenceof other disorders), the extent of bone loss in the individual, and thenutritional habits of the individual. Although the administration ofeven small quantities of calcium citrate malate and minerals may buildbone, the net increase in bone mass may not be detectable for shortperiods of administration.

For the treatment of age-related bone loss, the calcium citrate malateand minerals are administered for at least about six months, preferablyfor at least about twelve months. Of course, such administration may becontinued indefinitely, according to sound medical practice.

The methods of this invention may be employed in the treatment of any ofa variety of disorders in which the building of bone is desired. Thus,preferably, the human or other animal "subject" of the methods of thisinvention is "in need" of a method for building bone, i.e., the subjecthas a disorder for which building of bone or decrease in rate of boneresorption would be advantageous according to sound medical practice.Such disorders include, for example, bone fractures, broken mass anddisorders typified by bone loss, such as age-related bone loss andosteoporosis (both primary and secondary forms).

A preferred method of this invention is for the treatment of age-relatedbone loss. Such methods include administration of calcium citrate malatein combination with zinc, copper, manganese and other therapeuticagents. Estrogen therapy is commonly used. The method herein alsocomprises coadministering from about 0.6 mg to about 6 mg of estrogenalong with the calcium and trace minerals. Preferably from 0.625 mg toabout 1.25 mg of estrogen is taken daily. Any viable estrogen hormonereplacement can be used.

The following example illustrates compositions of the type provided bythe practice of this invention, but is not intended to be limitingthereof.

EXAMPLE I

Several post-menopausal women are treated by administering a compositioncontaining calcium citrate malate having molar calcium:citrate:malatecomposition of about 6:2:3. The calcium citrate malate is made by firstdissolving approximately 384.2 grams of citric acid and approximately402.3 grams of malic acid in approximately 2 liters of water. Thiscitrate/malate solution is then heated to approximately 55° C. (131°F.), with stirring. Separately, approximately 600.6 grams of calciumcarbonate is added to approximately 1.2 liters of water, forming aslurry, with stirring.

The citrate/malate solution is then removed from its heat source, andthe calcium carbonate slurry is added slowly, with stirring. The rate ofaddition is controlled, to contain the reaction as carbon dioxide isreleased. An additional quantity of water, approximately 0.4 liters, isfinally added. The reaction mixture is then stirred for approximately 1to 1.5 hours. The reaction is essentially complete as the pH of thesolution equilibrates to approximately 4.3.

A precipitate of calcium citrate malate is thus formed. The excessreaction liquid is filtered off. The calcium citrate malate is dried,for approximately 12 hours at approximately 105° C. (221° F.), reducingthe moisture level to less than about 1%. The dried product is thenmilled to approximately 10-20 mesh size, for a swallowable tabletformulation. Each tablet contains 250 mg.

The swallowable tablet dosage form is then made, comprising:

    ______________________________________                                        Component         % (By Weight)                                               ______________________________________                                        Calcium citrate malate*                                                                         99.73                                                       Magnesium stearate                                                                               0.27                                                       ______________________________________                                         *Having a molar calcium:citrate:malate composition of approximately 6:2:3     made as described above in this example.                                 

The tablet formulation is made by thoroughly admixing the powders, andtabletting using a standard tablet press, to form tablets weighingapproximately 1104 milligrams. The tablets are then coated, using a pancoater. The coating solution contains approximately 11%hydroxypropylmethyl cellulose, approximately 2% polyethylene glycol,approximately 3.5% colorant, and the balance of water.

A capsule containing 15 mg zinc (from zinc sulfate), 5 mg manganese(from manganese gluconate) and 2.5 mg copper (from copper gluconate) isalso administered to each patient.

The density of the subject's lumbar vertebrae is determined bydual-energy x-ray absorptiometry. The human subject is then administered4 of the calcium tablets and one trace mineral capsules comprised asabove, each day for 12 months. The mass of the subject's vertebrae isthen remeasured, indicating a net increase in bone mass. The followingresults are achieved after 1 year.

    ______________________________________                                        Change in Spine Bone Mineral Density                                                                             Net                                                      Number               Increase                                                 Of       % Change    In                                         Treatments    Patients After 1 Year                                                                              Bone Mass                                  ______________________________________                                        Calcium placebo/                                                                            42       -2.41 ± 0.63                                                                           0.00                                       Trace minerals placebo                                                        Calcium placebo/                                                                            38       -1.55 ± 0.66                                                                           0.86                                       trace minerals                                                                Calcium/trace minerals                                                                      34       -1.28 ± 0.70                                                                           1.13                                       placebo                                                                       Calcium and trace                                                                           37       -0.17 ± 0.67                                                                           2.24                                       minerals                                                                      ______________________________________                                    

The table shows that the calcium and trace minerals supplement affectsbone density and shows a net bone building effect of about 2.24%compared to the placebo group. The calcium trace mineral supplementeffect is significantly different from the dual placebo treatment at a95% confidence level. The dual placebo patients had at least a 2% lossover the one year period, the calcium/trace minerals patients showedlittle or no bone loss and thus a net increase of bone mass.

EXAMPLE II Preparation of Calcium-Citrate-Malate

A calcium-citrate-malate solution is prepared by dissolving 2 partssucrose and then 0.1 part citric and 0.28 part malic acids in 28.19parts water. Calcium hydroxide (0.22 part) is added and the mixture isagitated. This solution can be used directly to prepare beverages, orcan be freeze-dried to use in solid form.

EXAMPLE III Preparation of Calcium-Citrate-Malate Without Sugar

In an alternate mode, the sucrose can be deleted from the abovepreparation. Thus, a calcium citrate-malate solution is prepared byadmixing 62 g calcium carbonate with 11 g citric acid and 44 g malicacid dissolved in 1,040 g water at ambient temperature. This solutioncan be used to prepare low calorie beverages, beverage concentrates orfreeze-dried for use in solid supplements. The calcium is 53% of thesolid when dried to anhydrous conditions. Each ml of this solutionprovides 50 mg of calcium on an elemental basis.

EXAMPLE IV

A powdered mineral supplement comprising 2000 gm of calcium citratemalate prepared as above and 6.3 mg of copper sulfate (2.5 mg copper),31.3 mg of zinc chloride (15 mg zinc) and 5 mg of manganese (15.4 mg ofmanganese sulfate monohydrate) is prepared by tabletting the mixture ofpowders.

EXAMPLE V

Mineral calcium-fortified chewable lozenges comprise:

    ______________________________________                                        Ingredient             Amount                                                 ______________________________________                                        Calcium citrate-malate (6:2:3)                                                                       2500    mg                                             Dextrose               5       g                                              Fruit flavor*          6       mg                                             Zinc Chloride          31.3    mg                                             Manganese Gluconate    12.5    mg                                             Copper Gluconate       4.5     mg                                             Color                  As desired                                             ______________________________________                                         *Fruit flavors used herein generally comprise synthetically reconstituted     flavor esters. In this example, pineapple flavor is used and comprises a      synthetic mixture of ethyl acetate, acetaldehyde, methyl nvalerate, methy     .sub.- ivalerate, methyl .sub.- icaproate and methyl caprylate.          

The lozenge of Example IV is prepared by mixing the ingredients andcompacting the mixture in a standard press. This lozenge provides 936 mgcalcium, 15 mg zinc, 1.5 mg manganese and 1.43 mg copper.

EXAMPLE VI

A chocolate powder mix is prepared as follows:

    ______________________________________                                        Ingredient           Amount (percent)                                         ______________________________________                                        Granular Sucrose     66.88                                                    Non-fat Dry Milk     15.00                                                    Sodium Chloride      0.4                                                      Fermented Cocoa Powder, 14% fat                                                                    16.0                                                     Lecithin             1.0                                                      Colors               0.07                                                     Butylated Hydroxytoluene (BHT)                                                                     0.0003                                                   Ascorbic Acid        0.21                                                     Zinc Chloride        0.06                                                     Calcium Citrate Malate                                                                             0.05                                                     Artificial Chocolate Flavor                                                                        0.3                                                      ______________________________________                                    

The lecithin is heated to 45° C. to melt it. The cocoa is sterilized at160° C. in a pasteurizing oven for 2 hours.

A first premix is prepared by dry mixing the zinc salt and aboutone-half of the sodium chloride.

This premix is added to a dry mix of the remaining sodium chloride,vitamin C, and part of the non-fat dry milk. The colors and flavor arepremixed with part of the non-fat dry milk solids (0.8% of finalproduct). Cocoa powder, the remaining milk solids, minerals and thecolor/flavor mix are mixed together (22.88% of final product).

The BHT and lecithin are mixed together for one hour at 50° to 60° C.All of these mixes are then added to sucrose and blended to make ahomogeneous dry mix.

This beverage is taken along with a capsule containing 1.5 mg of zinc aszinc chloride, 5 mg of manganese as manganese gluconate, 2.0 mg ofcopper as cupric gluconate.

What is claimed is:
 1. A method for building of bone in a human subjectsuffering from age-related bone loss comprising administering to saidsubject a safe and effective amount of a mineral supplement comprisingcalcium citrate malate, zinc, manganese and copper salts for asufficient period of time to build bones in said subject, saidsupplement being in a liquid dosage form.
 2. A method for building ofbone according to claim 1, wherein said dosage is a beverage.
 3. Amethod according to claim 2 wherein said beverage contains juice.
 4. Amethod according to claim 3 wherein said juice is selected from thegroup consisting of orange juice, apple juice, pear juice and cranberryjuice.
 5. A method according to claim 1 wherein said mineral supplementis added to dry beverage mixes.
 6. A method for building of bone in ahuman subject suffering from age-related bone loss comprisingadministering to said subject a safe and effective amount of a mineralsupplement comprising calcium citrate malate, zinc, manganese and coppersalts for a sufficient period of time to build bone in said subject andwherein a safe and effective amount of estrogen is administered to thehuman subject.
 7. A method according to claim 6 wherein said estrogen isadministered in unit doses of from 0.6 mg to 6 mg.
 8. A method accordingto claim 7 wherein said estrogen dose is from 0.6 mg to 1.2 mg.
 9. Amethod of building bone according to claim 6, wherein said calciumcitrate malate is administered at a level of from about 175 mg to about2000 mg (on an elemental calcium basis), per day, said zinc isadministered at a level of 1.5 to 30 mg/day, said manganese at a levelof 1 to 8 mg/day and said copper is at a level of 1.0 to 5.0 mg/day.